LOCUS       V043679                10033 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043679
VERSION     V043679
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             234..414
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    458..583
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1076..1309
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1493..1537
                     /codon_start=1
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /product="antigenic peptide corresponding to amino acids
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
                     et al., 2013)"
                     /transl_table=1
                     /translation="KNEQELLELDKWASL"
                     /label="gp41 peptide"
     misc_feature    1804..1920
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2010..2213
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2302..4887
                     /label="Ldlr(NM_010700)"
                     /note="Ldlr(NM_010700)"
                     /gene="Ldlr"
     CDS             4894..4959
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        5003..5214
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             5227..5495
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5522..6121
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     misc_feature    6128..6716
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6790..7023
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    7095..7216
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7235..7370
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7408..7424)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7432..7448
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(7456..7486)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7501..7522
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7810..8398)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8569..9429)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(9430..9534)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     10008..10024
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt
     1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag
     2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct
     2281 agagctagcg aattcgccac catgagcacc gcggatctga tgcgtcgctg ggtcatcgcc
     2341 ctgctcctgg ctgctgccgg agttgcagca gaagactcat gcagcaggaa cgagttccag
     2401 tgtagagacg gaaaatgcat cgctagcaag tgggtgtgcg atggcagccc cgagtgcccg
     2461 gatggctccg atgagtcccc agagacatgc atgtctgtca cctgtcagtc caatcaattc
     2521 agctgtggag gccgtgtcag ccgatgcatt cctgactcct ggagatgtga tggacaggta
     2581 gactgtgaaa atgactcaga cgaacaaggc tgtcccccca agacgtgctc ccaggatgac
     2641 ttccgatgcc aggatggcaa gtgcatctcc ccgcagtttg tgtgtgatgg agaccgagat
     2701 tgcctagatg gctctgatga ggcccactgc caggccacca cttgtggccc cgcccacttc
     2761 cgctgcaact catccatatg catccccagt ctttgggcct gcgacgggga tgtcgactgt
     2821 gttgacggct ccgatgagtg gccacagaac tgccagggcc gagacacggc ctccaaaggc
     2881 gttagcagcc cctgctcctc cctggagttc cactgtggta gcagtgagtg tatccatcgc
     2941 agctgggtct gtgacggcga ggcagactgc aaggacaagt cagatgagga gcactgcgcg
     3001 gtggccacct gccgacctga tgaattccag tgtgcagatg gctcctgcat tcacggtagc
     3061 cgccagtgtg accgtgaaca tgactgcaag gacatgagcg acgagctcgg ctgcgtcaat
     3121 gtgacacagt gtgatggccc caacaagttc aagtgtcaca gtggggagtg catcagcttg
     3181 gacaaggtgt gcgactccgc ccgcgactgc caggactggt cggatgagcc catcaaggag
     3241 tgcaagacca acgagtgttt ggacaacaat ggtggctgtt cccacatctg caaggacctc
     3301 aagattggct ctgagtgcct gtgtcccagc ggcttccggt tggtggacct ccacaggtgt
     3361 gaagatattg acgagtgtca ggagccagac acctgcagcc agctctgtgt gaacctggaa
     3421 ggcagctaca agtgtgagtg ccaggccggc ttccacatgg acccacacac cagggtctgc
     3481 aaggctgtgg gctccatagg ctatctgctc ttcaccaacc gccacgaggt ccggaagatg
     3541 accctggacc gcagcgagta caccagtctg ctccccaacc tgaagaatgt ggtggctctc
     3601 gacacggagg tgaccaacaa tagaatctac tggtccgacc tgtcccaaaa aaagatctac
     3661 agcgccctga tggaccaggc ccctaacttg tcctacgaca ccatcatcag tgaggacctg
     3721 catgcccctg acgggctggc ggtagactgg atccaccgca acatctactg gacagattca
     3781 gtcccaggca gcgtatctgt ggctgacacc aagggcgtaa agaggaggac actgttccaa
     3841 gaggcagggt ccagacccag agccatcgta gtggaccctg tgcatggctt catgtactgg
     3901 acagattggg gaacacccgc caagatcaag aaagggggtt tgaatggtgt ggacatccac
     3961 tcactggtga ccgaaaacat ccagtggcca aatggcatca cactagatct ttccagtggc
     4021 cgtctctatt gggttgattc caaactccac tctatctcca gcatcgatgt caatgggggc
     4081 aatcggaaaa ccattttgga ggatgagaac cggctggccc accccttctc cttggccatc
     4141 tatgaggaca aagtgtattg gacagatgtc ataaacgaag ccattttcag tgccaatcga
     4201 ctcacgggtt cagatgtgaa tttggtggct gaaaacctct tgtccccgga ggacattgtc
     4261 ctgttccaca aggtcacaca gcctagaggg gtgaactggt gtgagacaac agccctcctc
     4321 cccaatggtg gttgccagta cctgtgcctg cccgccccac agatcggtcc ccactcgccc
     4381 aaattcacct gcgcctgccc tgatggcatg ctgctggcca aggacatgcg gagctgcctc
     4441 acagaagtcg acactgtact gaccacccag gggacatccg ccgtccggcc tgtggtcacc
     4501 gcatcagcta ccaggccacc gaagcacagt gaggatctct cagctcccag tactcctagg
     4561 cagcctgtgg acaccccagg gctcagcaca gtggcgtcag tgacagtgtc ccaccaagtc
     4621 cagggtgaca tggctggcag agggaatgag gagcagccac atggtatgag gttcctgtcc
     4681 atcttcttcc ctattgcact ggttgccctc cttgtccttg gggccgtcct gctgtggagg
     4741 aactggcggc tgaagaacat caacagcata aactttgaca acccagtcta ccagaagacc
     4801 acagaggacg agctccacat ttgccgaagc caggatggct atacctaccc ctcaagacag
     4861 atggtcagcc tggaggacga tgtggcagga tccgactaca aagaccatga cggtgattat
     4921 aaagatcatg acatcgacta caaggatgac gatgacaagt agcggccgcg aaggatctgc
     4981 gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg
     5041 gggggagggg tcggcaattg aacgggtgcc tagagaaggt ggcgcggggt aaactgggaa
     5101 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt
     5161 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acagctgaag
     5221 cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc ctacctgagg ccgccatcca
     5281 cgccggttga gtcgcgttct gccgcctccc gcctgtggtg cctcctgaac tgcgtccgcc
     5341 gtctaggtaa gtttaaagct caggtcgaga ccgggccttt gtccggcgct cccttggagc
     5401 ctacctagac tcagccggct ctccacgctt tgcctgaccc tgcttgctca actctacgtc
     5461 tttgtttcgt tttctgttct gcgccgttac agatccaagc tgtgaccggc gcctacgcta
     5521 gatgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt
     5581 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgacccgga
     5641 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
     5701 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
     5761 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
     5821 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
     5881 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg
     5941 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
     6001 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
     6061 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg
     6121 agtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     6181 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     6241 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     6301 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     6361 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     6421 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     6481 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     6541 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     6601 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     6661 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg
     6721 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
     6781 aggggggact ggaagggcta attcactccc aacgaaaata agatctgctt tttgcttgta
     6841 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     6901 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     6961 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     7021 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat
     7081 cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     7141 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     7201 tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc cgcccagttc
     7261 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     7321 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc tagacttttg
     7381 cagagacggc ccaaattcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     7441 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     7501 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     7561 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     7621 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     7681 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     7741 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     7801 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     7861 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     7921 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     7981 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     8041 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     8101 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     8161 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     8221 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     8281 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     8341 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     8401 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     8461 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     8521 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     8581 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     8641 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     8701 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     8761 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     8821 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     8881 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     8941 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     9001 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     9061 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     9121 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     9181 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     9241 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     9301 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     9361 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     9421 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     9481 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     9541 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     9601 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     9661 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     9721 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     9781 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     9841 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga
     9901 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     9961 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
    10021 cagtgccaag ctg
//
